A High-Throughput Assay for Collagen Secretion Suggests an Unanticipated Role for Hsp90 in Collagen Production

Madeline Y. Wong, Ngoc Duc Doan, Andrew S. Dichiara, Louis J. Papa, Jaime H. Cheah, Christian K. Soule, Nicki Watson, John D. Hulleman, Matthew D. Shoulders

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

Collagen overproduction is a feature of fibrosis and cancer, while insufficient deposition of functional collagen molecules and/or the secretion of malformed collagen is common in genetic disorders like osteogenesis imperfecta. Collagen secretion is an appealing therapeutic target in these and other diseases, as secretion directly connects intracellular biosynthesis to collagen deposition and biological function in the extracellular matrix. However, small molecule and biological methods to tune collagen secretion are severely lacking. Their discovery could prove useful not only in the treatment of disease, but also in providing tools for better elucidating mechanisms of collagen biosynthesis. We developed a cell-based, high-throughput luminescent assay of collagen type I secretion and used it to screen for small molecules that selectively enhance or inhibit that process. Among several validated hits, the Hsp90 inhibitor 17-allylaminogeldanamycin (17-AAG) robustly decreases the secretion of collagen-I by our model cell line and by human primary cells. In these systems, 17-AAG and other pan-isoform Hsp90 inhibitors reduce collagen-I secretion post-translationally and are not global inhibitors of protein secretion. Surprisingly, the consequences of Hsp90 inhibitors cannot be attributed to inhibition of the endoplasmic reticulum's Hsp90 isoform, Grp94. Instead, collagen-I secretion likely depends on the activity of cytosolic Hsp90 chaperones, even though such chaperones cannot directly engage nascent collagen molecules. Our results highlight the value of a cell-based high-throughput screen for selective modulators of collagen secretion and suggest an unanticipated role for cytosolic Hsp90 in collagen secretion.

Original languageEnglish (US)
Pages (from-to)2814-2827
Number of pages14
JournalBiochemistry
Volume57
Issue number19
DOIs
StatePublished - May 15 2018

ASJC Scopus subject areas

  • Biochemistry

Fingerprint Dive into the research topics of 'A High-Throughput Assay for Collagen Secretion Suggests an Unanticipated Role for Hsp90 in Collagen Production'. Together they form a unique fingerprint.

  • Cite this

    Wong, M. Y., Doan, N. D., Dichiara, A. S., Papa, L. J., Cheah, J. H., Soule, C. K., Watson, N., Hulleman, J. D., & Shoulders, M. D. (2018). A High-Throughput Assay for Collagen Secretion Suggests an Unanticipated Role for Hsp90 in Collagen Production. Biochemistry, 57(19), 2814-2827. https://doi.org/10.1021/acs.biochem.8b00378